Fig. 2: Pre-rRNA expression is inversely correlated with lifespan and healthspan.

a, b RT-qPCR analysis of ncl-1 mRNA (a) and pre-rRNA (b) levels of worms treated either with control (Ctrl) RNAi or ncl-1 RNAi. Values are relative to Ctrl RNAi and normalized to tgb-1 mRNA. Mean ± S.D. values of 3 independent experiments (n = 60 worms per sample) are shown. c Lifespan analysis of the nematodes upon treatment with Ctrl RNAi or ncl-1 RNAi, n = 140 worms. Survival was scored daily and P values were calculated using the Mantel-Cox test. Result is representative of 3 independent trials. d Body size (area) measurement of animals treated with Ctrl, tif-1A or ncl-1 RNAi. n = 30 worms. Data combine three independent trials and mean ± S.D. is shown. e Schematic of the locomotor performance test (burrowing assay). f, g Burrowing assays of animals treated with Ctrl, tif-1A or ncl-1 RNAi. The % of animals reaching the surface was determined at the indicated time points. Data show mean values ± S.D. of 3 independent assays performed on the second day of adulthood (AD2) with n = 40–45 worms (f) or on AD12 with n = 40–60 worms (g). P values were calculated with the Mantel-Cox test. h, i Representative images (n = 10 worms per image) and quantification of intestinal integrity monitored by the Brilliant Blue dye retention assay. Worms with staining outside the gut (‘Smurf’ phenotype) were scored following treatment with Ctrl, tif-1A or ncl-1 RNAi (n = 40–60 worms per treatment) at AD2 (h) or AD12 (i), scale bar 200 μm. Mean ± S.D. values of 3 independent experiments are shown. P values were determined using a Student’s unpaired t-test. ns = not significant, ****P < 0.0001, ***P < 0.001, **P < 0.01, *P < 0.05. All statistical tests used were two-sided, the exact P values and statistical analyses are reported in the Source Data file.